The specific goal of this developmental project is to optimize and characterize conformation-specific single-chain Fv (scFv) antibodies as tools to recognize and manipulate misfolded neurodegeneration proteins intracellularly. We have developed unique capabilities that enable us to generate scFvs that target specific protein morphologies and to test their function intracellularly. Given the known structural commonality of proteins involved in Alzheimer's, Parkinson's, Huntington's, prion, and ALS diseases, these reagents will allow investigators working on these various diseases to distinguish between oligomeric and fibrillar forms of aggregates in a cellular context. It will also allow redirection of specific forms of proteins to other intracellular compartments to test hypotheses of where the most critical site of action resides. This is a multiple-PI application. The anti-alpha-synuclein oligomeric and anti- fibrillar scFvs are currently being selected in the laboratory of Dr. Michael Sierks, at ASU. Much of the first year will be devoted to efficient rapid screening of intracellular expression in neuronal cell lines by Dr. Anne Messer, Wadsworth Center in Albany, NY, followed immediately by the more labor-intensive engineering of affinity and stability of the most promising scFvs. By the second year, the majority of the work will consist of detailed biological studies done in Albany, with further protein interactions and optimization as needed at ASU. PUBLIC HEALTH REVELANCE: Neurodegenerative diseases are taking an increasing human and financial toll as our population ages. Engineered antibody therapeutics are already in use for cancer and infectious disease, and offer an exciting new platform for neurological disease drug discovery and therapeutics. This project specifically targets the consequences of protein misfolding in Parkinson's disease, with direct applications to related diseases such as Alzheimer's, Huntington's, and ALS.